Selection of reference genes for qPCR normalization in buffalobur (Solanum rostratum Dunal)
Selection of reference genes for qPCR normalization in buffalobur (Solanum rostratum Dunal)
复制标题
选择水牛 (Solanum rostratum Dunal) qPCR 标准化参考基因
DOI：
10.1038/s41598-019-43438-6
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发表时间：
2019-05-06
影响因子：
4.6
通讯作者：
Wei, Shouhui
中科院分区：
文献类型：
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作者：
Zhao, Dandan;Wang, Xu;Wei, Shouhui
水牛(Solanum RostratumDunal)隶属茄科，是一种世界性的外来入侵杂草，被《中国》列为十大外来入侵物种之一。它对人类和牲畜都是有害的，因为整个植物上长满了含有毒素的脊椎。许多研究已经利用定量实时聚合酶链式反应(QPCR)分析了该杂草在不同胁迫条件下的基因表达。然而，到目前为止，还没有关于水牛合适的参考基因的报道。本研究选择了14个候选参考基因，并利用geNorm、NormFinder、BestKeeper和RefFinder统计算法对其在水牛不同组织、不同发育阶段以及对几种胁迫条件下的表达稳定性进行了评估。结果表明，EF1基因α、ACT和SAND在所有供试样品中都是合适的参考基因。我们建议在不同的实验条件下联合使用这三个基因来标准化靶基因的表达。通过评估P5CSI和GI的相对表达水平，对所选参考基因进行验证。这项工作为水牛的转录标准化找到了合适的参考基因，这将有助于今后对这种入侵杂草的遗传学研究。
Buffalobur (Solanum rostratumDunal), which belongs to the Solanaceae family, is a worldwide noxious invasive weed and is listed as one of the top 10 alien invasive species in China. It is harmful to humans and livestock because the entire plant is covered with spines containing toxins. Many studies have analysed the gene expression in this weed species under different stress conditions using quantitative real-time PCR (qPCR). However, until now, there has been no report on suitable reference genes in buffalobur. Herein, 14 candidate reference genes were selected and evaluated for their expression stability in buffalobur in different tissues, at different developmental stages, and in response to several stress conditions using the geNorm, NormFinder, BestKeeper and RefFinder statistical algorithms. The results showed thatEF1α,ACTandSANDare suitable reference genes across all samples tested. We recommend the normalization of target gene expression under different experimental conditions using these three genes together. Validation of selected reference genes was achieved by assessing the relative expression levels ofP5CSandGI. This work identified the appropriate reference genes for transcript normalization in buffalobur, which will be helpful in future genetic studies of this invasive weed.